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A Novel Amperometric Glutamate Biosensor Based on Glutamate Oxidase Adsorbed on Silicalite

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ABSTRACT

In this work, we developed a new amperometric biosensor for glutamate detection using a typical method of glutamate oxidase (GlOx) immobilization via adsorption on silicalite particles. The disc platinum electrode (d = 0.4 mm) was used as the amperometric sensor. The procedure of biosensor preparation was optimized. The main parameters of modifying amperometric transducers with a silicalite layer were determined along with the procedure of GlOx adsorption on this layer. The biosensors based on GlOx adsorbed on silicalite demonstrated high sensitivity to glutamate. The linear range of detection was from 2.5 to 450 μM, and the limit of glutamate detection was 1 μM. It was shown that the proposed biosensors were characterized by good response reproducibility during hours of continuous work and operational stability for several days. The developed biosensors could be applied for determination of glutamate in real samples.

No MeSH data available.


Scheme of platinum disc electrodes. 1 Bioselective membrane, 2 platinum wire, 3 inner conductor, 4 wood alloy, 5 epoxy resin, and 6 contact panel
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Fig2: Scheme of platinum disc electrodes. 1 Bioselective membrane, 2 platinum wire, 3 inner conductor, 4 wood alloy, 5 epoxy resin, and 6 contact panel

Mentions: In this work, the platinum disc electrodes of own production served as amperometric transducers (Fig. 2). A platinum wire of 0.4 mm in diameter and 3 mm long was sealed in the end of a glass capillary with an outer diameter of 3.5 mm. An open end of the wire served as the transducers working surface. An inner end of the platinum wire was connected to a copper wire, placed inside the capillary, using fusible Wood’s alloy. A contact pad for connecting the electrode to the measuring setup was placed at the other end of the copper wire. The working surface of platinum electrodes was cleaned by immersing it into a concentrated sulfuric acid for 30 s, washing with water and ethanol-wetted cotton swab prior to immobilization of the bioselective element. The surface of working electrode was obtained by grinding with alumina powder (particles of 0.1 and 0.05 μm) and treated with alcohol before the bioselective element immobilization. The electrode surface was periodically restored using the same grinding procedure.Fig. 2


A Novel Amperometric Glutamate Biosensor Based on Glutamate Oxidase Adsorbed on Silicalite
Scheme of platinum disc electrodes. 1 Bioselective membrane, 2 platinum wire, 3 inner conductor, 4 wood alloy, 5 epoxy resin, and 6 contact panel
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5383914&req=5

Fig2: Scheme of platinum disc electrodes. 1 Bioselective membrane, 2 platinum wire, 3 inner conductor, 4 wood alloy, 5 epoxy resin, and 6 contact panel
Mentions: In this work, the platinum disc electrodes of own production served as amperometric transducers (Fig. 2). A platinum wire of 0.4 mm in diameter and 3 mm long was sealed in the end of a glass capillary with an outer diameter of 3.5 mm. An open end of the wire served as the transducers working surface. An inner end of the platinum wire was connected to a copper wire, placed inside the capillary, using fusible Wood’s alloy. A contact pad for connecting the electrode to the measuring setup was placed at the other end of the copper wire. The working surface of platinum electrodes was cleaned by immersing it into a concentrated sulfuric acid for 30 s, washing with water and ethanol-wetted cotton swab prior to immobilization of the bioselective element. The surface of working electrode was obtained by grinding with alumina powder (particles of 0.1 and 0.05 μm) and treated with alcohol before the bioselective element immobilization. The electrode surface was periodically restored using the same grinding procedure.Fig. 2

View Article: PubMed Central - PubMed

ABSTRACT

In this work, we developed a new amperometric biosensor for glutamate detection using a typical method of glutamate oxidase (GlOx) immobilization via adsorption on silicalite particles. The disc platinum electrode (d = 0.4 mm) was used as the amperometric sensor. The procedure of biosensor preparation was optimized. The main parameters of modifying amperometric transducers with a silicalite layer were determined along with the procedure of GlOx adsorption on this layer. The biosensors based on GlOx adsorbed on silicalite demonstrated high sensitivity to glutamate. The linear range of detection was from 2.5 to 450 μM, and the limit of glutamate detection was 1 μM. It was shown that the proposed biosensors were characterized by good response reproducibility during hours of continuous work and operational stability for several days. The developed biosensors could be applied for determination of glutamate in real samples.

No MeSH data available.